EP1412421A1 - Polymere biocompatible a structure tridimensionnelle a cellules communicantes, procede de preparation et application en medecine et en chirurgie - Google Patents
Polymere biocompatible a structure tridimensionnelle a cellules communicantes, procede de preparation et application en medecine et en chirurgieInfo
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- EP1412421A1 EP1412421A1 EP01949519A EP01949519A EP1412421A1 EP 1412421 A1 EP1412421 A1 EP 1412421A1 EP 01949519 A EP01949519 A EP 01949519A EP 01949519 A EP01949519 A EP 01949519A EP 1412421 A1 EP1412421 A1 EP 1412421A1
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- polymer
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- hydrogel
- soluble
- cells
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
- A61L27/18—Macromolecular materials obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
- A61L27/16—Macromolecular materials obtained by reactions only involving carbon-to-carbon unsaturated bonds
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/38—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells
- A61L27/3804—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells characterised by specific cells or progenitors thereof, e.g. fibroblasts, connective tissue cells, kidney cells
- A61L27/3817—Cartilage-forming cells, e.g. pre-chondrocytes
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/38—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells
- A61L27/3839—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells characterised by the site of application in the body
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/52—Hydrogels or hydrocolloids
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/54—Biologically active materials, e.g. therapeutic substances
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/56—Porous materials, e.g. foams or sponges
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J9/00—Working-up of macromolecular substances to porous or cellular articles or materials; After-treatment thereof
- C08J9/26—Working-up of macromolecular substances to porous or cellular articles or materials; After-treatment thereof by elimination of a solid phase from a macromolecular composition or article, e.g. leaching out
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/20—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
- A61L2300/258—Genetic materials, DNA, RNA, genes, vectors, e.g. plasmids
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/60—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a special physical form
- A61L2300/64—Animal cells
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2323/00—Characterised by the use of homopolymers or copolymers of unsaturated aliphatic hydrocarbons having only one carbon-to-carbon double bond; Derivatives of such polymers
- C08J2323/02—Characterised by the use of homopolymers or copolymers of unsaturated aliphatic hydrocarbons having only one carbon-to-carbon double bond; Derivatives of such polymers not modified by chemical after treatment
- C08J2323/18—Homopolymers or copolymers of hydrocarbons having four or more carbon atoms
- C08J2323/20—Homopolymers or copolymers of hydrocarbons having four or more carbon atoms having four to nine carbon atoms
Definitions
- the present invention relates to the field of bio-materials. It relates more particularly to a process for the preparation of a biocompatible porous polymer with communicating cavities of controlled size, porosity and rigidity. It relates in particular to the application of these biocompatible materials to the culture of cells in vitro, as well as to the preparation of biocompatible supports thus seeded, as they are or encapsulated by a polymer or its semi-permeable and also biocompatible hydrogel, as implants in different organs or tissues of the human or animal body, to permanently or temporarily replace a failing organ, and thus create a bio-artificial organ. This could be the case of the bio-artificial pancreas, bio-artificial liver, bio-artificial cornea, bio-artificial articular cartilage, bio-artificial bone, etc.
- tissue or cell growth factor more generally a biologically active substance such as a cytokine, a growth factor or a recombinant molecule of therapeutic interest.
- This porous material with communicating cavities can also be implanted "naked" in the living body, to fill the deficit of substances such as for example: the cartilaginous substance in maxillofacial surgery or the production of breast prostheses.
- Biomaterials according to the invention also find their application in the preparation of filters for bio-purification of biological fluids, or as carriers of enzymes for the production of an enzymatic reactor.
- bio-materials must be understood as non-living materials used in a medical device intended to interact with biological systems.
- a bio-material is suitable for contact with living tissues or fluids or living tissues. This contact, which is evident in the case of an implant, must be extended to the contact which takes place on the surface or outside of the human or animal body, for example those which occur with blood in hemodialysis or with the cornea in contact lenses. It is also extended to materials which can be used in biotechnology, in particular materials for the in vitro culture of animal or plant cells.
- a bio-material is in essence biocompatible.
- bio-compatibility is meant the ability of a material to be used with an appropriate host response in a specific application.
- This definition implies “negative” properties such as the absence of toxicity, the absence of inflammatory reaction, the absence of activation of the complement, the absence of leukocyte fall.
- This also includes “positive properties” which imply that the material is not necessarily as inert as possible but on the contrary makes react the living tissue and contributes to the metabolic activation of the cells which are in contact with it or the tissues in which it is implanted; this is particularly the case with osteoconductive materials which facilitate bone growth.
- the porous bio-materials of the present invention fall into the category of organic polymers or copolymers.
- the method of the invention and the material with communicating cavities obtained by the method allow cells placed in culture to organize in these communicating cavities and if necessary proliferate there.
- These properties allow not only the cultivation of cells and the manufacture of artificial organs, but also the construction of transportable and transplantable cellular tissues, in particular in the transplantation of neocartilaginous tissues, produced from chondrocytes in culture.
- Various porous materials based on natural or synthetic, organic or mineral products have been described in their use in the culture of cells in vitro and in the transplantation of living cells.
- hydroxy apatite PAH
- TCB tricalcium phosphate- ⁇
- the phosphocalcic hydroxy apatite of formula Ca-to (PO4) 6 OH 2 is a synthetic material sold as a synthetic bone substitute by the company TECHNIMED.
- the difficulty with this type of material is to manage to synthesize a PAH having just the right size of pores so that colonization by cells, and in particular bone cells, is done correctly.
- the use of these types of materials is limited by the lack of knowledge of the degradation mechanisms, their durability and resistance to fracture, their surface activity and the possibilities of calcification.
- polyester or polytetrafluoroethylene felts alone and treated with a polyurethane (1); polyethyl methacrylate / tetrahydrofurfuryl methacrylate (2) and (3), collagen sponges (4); polyhydroxyethylmetacrylate (5); copolymers of polyglycolic and polylactic acids (6).
- Shapiro L. and Cohen S. prepared a rigid alginate sponge, intended to be seeded by cells, followed by cultivation and transplantation into the living body.
- the various applications also require the implementation of a process making it possible to control the size of the cavities, the shape and the rigidity (the more or less great flexibility) of the polymer.
- these objectives can be achieved with a copolymer of the family of a polymer used for many years in the form of membranes for hemodialysis or in its hydrogel form, for ocular implants or for the preparation of pancreas. artificial (7).
- This copolymer has already been shown to be biocompatible and hemo-compatible, and in particular as regards its capacities not to activate the complement system (15), not to induce a leukocyte fall and to induce only a minimal hypoxemia (8 ).
- the polymer in question is a copolymer called AN 69, manufactured by the company HOSPAL R. & D. Int.
- the process of the invention uses the very properties of the manufacture of the hydrogel illustrated in the case of the copolymer of acrylonitrile and sodium methallylsulfonate, said manufacture successively comprising a solution step and a gelling step then of forming a hydrogel .
- the formation and definition of hydrogels is described by Honiger et al. (7).
- the hydrogel is formed by precipitation of a homogeneous polymer solution. In a ternary diagram (polymer / solvent / non-solvent), the equilibrium curve separates an area where all the components are miscible from another area or two phases are formed (a solid phase rich in polymer and a liquid phase poor or depleted in polymer).
- the system evolves from the initial polymer solution to a composition where all the solvent is replaced by the non-solvent, this transforming the gel into hydrogel; this hydrogel essentially only contains non-solvent and polymer.
- This succession of steps (liquid form, gelled form), the transition from the liquid form to the gelled form being triggered by contact of the copolymer with a non-solvent, makes it possible to envisage carrying out this gelling around a form matrix. and of porosity previously chosen as a function of the subsequent application of the biocompatible copolymer.
- the concept underlying the invention is to use a mold or a matrix which gives the biomaterial the chosen porosity and shape, the rigidity being determined by the conditions for producing the hydrogel and in particular its content. in water.
- the process resides on an essential characteristic of the polymer which is the capacity to pass from a liquid state to a non-liquid state, having a certain rigidity.
- the present invention applies by equivalence to any biocompatible polymer which can, thanks to a triggering factor, pass from a liquid state to a non-liquid state.
- non-liquid state is meant a gelled or crystallized or pseudo-crystallized state, or a hydrogel state.
- the choice of mold or matrix used to give the bio-material its shape and porosity is made according to two alternative strategies.
- the first is to choose a material for the mold or the matrix whose neutrality and bio-compatibility are total. But as we said above, today no material is known which has at the same time the capacities to have a controlled porosity, a bio- compatibility and control of these long-term effects.
- the second alternative is to use as a mold or matrix any substance whose size and porosity are controllable and capable, after the formation of the hydrogel or of the solid structure on said mold, of being eliminated. This elimination can be carried out by dissolution, or by enzymatic digestion.
- a first embodiment of the invention relates to a process for obtaining a porous three-dimensional structure with communicating cavities consisting of a biocompatible polymer comprising a liquid state and a gelled or solid state, comprising the following operations: - preparing a frit of selected geometry and porosity consisting of a water-soluble or hydrolyzable substance, not soluble in the solvent for the polymer and soluble in the non-solvent for the polymer,
- the frit by immersing the mixture in a non-solvent for the polymer, - recover the polymer of geometry and porosity chosen in gelled or solid form or in the form of a hydrogel.
- the invention relates to a process for obtaining a three-dimensional porous structure with communicating cavities consisting of a biocompatible polymer comprising a liquid state and a gelled or solid state, comprising the operations following:
- the mold intended to give its shape and its dimension to the three-dimensional structure may for example be made of silicone elastomer.
- the two embodiments of the process described above are based on two essential characteristics: a) the biocompatible polymer used must have the properties of passing from a liquid state to a gelled or solid state, this transformation being able to be controlled by an external trigger non-solvent type of polymer, temperature, pH, for example, i.e. the property of forming a hydrogel; b) the use of a matrix which performs the role of a mold intended to give the desired shape to the biocompatible polymer, said matrix or said mold being able to be eliminated, either by dissolution, or by hydrolysis.
- the external shape which can be chosen according to a desired geometry for an implant.
- the production of an implant for bone regeneration must have the desired geometry for perfect implantation in an insertion site.
- the form can thus be produced, either by conferring from the outset a form on the matrix consisting of the water-soluble or hydrolysable substance, this constituting the first embodiment of the process, or by preparing the water-soluble or hydrolysable substance in the form of beads or microbeads whose size is chosen according to the desired size for the communicating cavities; these beads or these microbeads are then mixed with the liquid form of the biocompatible polymer, said mixture being prepared in a mold of selected geometry and size. After gelation and solidification of the polymer, the mold is then removed before or after dissolution or hydrolysis of the substance.
- the gelation or solidification step is carried out by immersion in a bath containing a non-solvent for the polymer.
- the bath known as gelling or solidifying bath or of hydrogel formation comprises water or an aqueous solution of a biologically acceptable salt.
- the biomaterials in question fall into the category of hydrogels.
- the hydrogels are three-dimensional hydrophilic networks that are capable of absorbing large amounts of water or biological fluid and that to some extent resemble biological tissue. They are insoluble due to the presence of a network of chemical or physical bonds, and can be formed in response to a large number of physiological or physical stimuli such as temperature, ionic strength, pH or contact with solvents.
- the three-dimensional structures are essentially based on a hydrogel, that is to say that the structure is made of a homogeneous material.
- the polymer solution comprises at least:
- aprotic solvent any solvent which does not exchange protons with the ambient medium or the substances dissolved in it.
- a preferred hydrogel contains 50 to 98% water.
- the ionic capacity of the hydrogel can be between 0 and approximately 500 mEq / kg, preferably 30 to 300 mEq / kg, more preferably between 100 and 270 mEq / kg of hydrogel. Low ionic capacities (of the order of 0) are reached for the homopolymer hydrogel PAN (AN69 without sodium methallylsulfonate group).
- Such hydrogels can be formed from a solution of polymers comprising at least: - a copolymer of acrylonitrile and of an unsaturated olefinic comonomer carrying anionic groups, said comonomer being chosen in the group consisting of metallylsulfonic acid, metallylcarboxylic acid, metallylphosphoric acid, metallylphosphonic acid, optionally salified metallylsulfiric acid,
- the polymer solution in the solvent additionally contains non-solvent for the polymer.
- the copolymer is a copolymer of acrylonitrile and sodium metallylesulfonate.
- This polymer has been described and used as a biocompatible material in numerous applications.
- This polymer is AN 69, which is referred to above. It is a copolymer of sodium polyacrylonitrile-methallylsulfolnate with a molecular weight of approximately 250,000. Its anionic character depends on the content of the sulfonic group (3.3 mol%).
- This copolymer can be dissolved in an aprotic solvent such as
- NN-dimethylformamide (DMF), dimethylsulfoxide (DMSO), NN-dimethylacetamide (DMAA), or propylene carbonate (PC).
- DMF dimethylsulfoxide
- DMAA dimethylsulfoxide
- PC propylene carbonate
- the polymer can also be chosen from a group containing polysulfone, polyethersulfone, polyhydroxy ethyl methacrylate, polyhydroxy propyl methacrylate, or their copolymers.
- the hydrogel may contain from 2 to 50% of acrylonitrile copolymers and of an unsaturated comonomer carrying anionic groups, the acrylonitrile / co-monomer molar ratio being between 90:10 and 100: 0 .
- Such a hydrogel has a micro-porous structure and an ionic capacity of between 0 and 500 mEq per kilo of gel, a water content of between 50 and 98%.
- This polymer has been used for more than twenty years as a renal dialysis membrane in the form of hollow fiber or flat sheet. Its physical and chemical properties are well known, and it has proven for over twenty years its excellent bio-compatibility with blood and serum. In particular, as early as 1978, it was established that the membrane of AN 69 did not generate activation of the complement giving an aggregation of leukocytes nor a sequestration of the aggregates formed in the pulmonary microcirculation leading in turn to a leukopenia and at risk of hypoxia (11).
- the aprotic solvent for the copolymer will preferably be, when the polymer is a copolymer of acrylonitrile and of a metallylesulfonate comonomer, chosen from a group comprising NN-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), NN-dimethylacetamide, polypropylene carbonate, N-methylpyrrolidone (NMP).
- a metallylesulfonate comonomer chosen from a group comprising NN-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), NN-dimethylacetamide, polypropylene carbonate, N-methylpyrrolidone (NMP).
- DMF NN-dimethylformamide
- DMSO dimethyl sulfoxide
- NMP N-methylpyrrolidone
- each of the elements making up the polymer solution can vary according to the characteristics expected for the biocompatible polymer, in particular as regards its rigidity.
- a material according to the invention comprising 5 to 15% of polymer will give a flexible, deformable sponge.
- a material containing 25 to 35% of polymer will be preferred and makes it possible to obtain a porous substance of controlled rigidity / flexibility, as a function of the weight ratio of the polymer or copolymer on the one hand, and of the water-soluble or hydrolyzable substance, on the other hand.
- a geometry or porosity frit prepared with a water-soluble or hydrolyzable substance, or this same substance prepared in the form of particles of size and chosen geometry is soaked or mixed with the biocompatible polymer in its liquid state.
- the methods of the invention are carried out essentially without evaporation of the solvent or of the non-solvent.
- the water-soluble or hydrolyzable substance which is not soluble in the polymer solvent and soluble in the polymer non-solvent is agglomerated or crystallized sucrose.
- this substance may be an agglomerate of cane or beet sugar pseudo-crystals in pieces or in powder.
- the advantages of using this substance are its perfect tolerability in terms of toxicity, its very good solubility and finally the ease with which one can modulate the shape and size of the agglomerated particles.
- sucrose makes it possible to obtain particles with an average diameter of between 0.1 and 3 mm, thus giving the biocompatible polymer communicating cavities of the chosen size. In certain cases, after elimination of the sucrose, a certain withdrawal of the cavities can appear. This is homogeneous within the foam obtained and reproducible for a given polymer or copolymer.
- a person skilled in the art will be able to choose the size of the particles as a function of the size of the communicating cavities sought.
- the non-solvent for the polymer is an aqueous solution of an organic or inorganic salt.
- the polymer solution is composed of copolymers of acrylonitriles and sodium metallylesulfonate, the aprotic solvent being DMSO
- the non-solvent of said polymer capable of forming the hydrogel is a chloride solution.
- the water-soluble or hydrolyzable substance is, if necessary, eliminated by immersion in distilled water at a temperature between 30 and 50 °, preferably with stirring. The water is renewed until the sugar crystals are completely dissolved, and the communicating cavities are released.
- the average diameter of the cavities can be between 0.1 and 3 mm.
- the diameter of the cavities depends of course on the size of the particles of the water-soluble or hydrolyzable substance which are eliminated, while being able to be smaller, because of a withdrawal effect observed during the formation of the hydrogel.
- the present invention also relates to a porous three-dimensional structure with communicating cavities consisting of at least one biocompatible polymer, capable of being obtained by a process as described above; these structures have multiple cavities, which communicate with each other and with the surface of said structure.
- This three-dimensional, porous hydrogel structure with communicating cells can be described as "foam”.
- the term "foam” describes both the existence of cavities communicating with each other and with the surface of said foam, as the fact of having rigidities and variable geometries.
- the term "foam” or "polymer foam” will designate all the three-dimensional structures capable of being obtained by the process of the invention.
- the polymer foams of the invention will be hydrogel foams and more preferably AN 69 foams obtained by the process.
- the hydrogel foams according to the invention, and more particularly the AN 69 foams can contain functionalized residues capable of forming covalent bonds with organic residues.
- these functionalized residues can be residues -CHO, -NH 2 , -COOH, -SH.
- An example of such a functionalization is described in patent application PCT / FR 98/00066 for the AN 69. This example is nonetheless non-limiting insofar as in patent applications WO 92/07023 and WO 92/07006, it has also been described the functionalization of other uncharged hydrophilic polymers such as polyethylene glycol-hypoxy covalently linked to a polyethyleneimine.
- the advantage of the foams according to the invention carrying functionalized residues is the possibility of coupling, by covalent or ionic bond, organic ligands; by way of example, such ligands can be selected from a group comprising antibodies, antigens, peptides, proteins or glycoproteins, hormones, enzymes, co-factors thereof, substrates or inhibitors of these, polysaccharides, lectins, toxins or anti-toxins, nucleic acids or polynucleotides, haptens or hapten ligands, pigments or dyes.
- organic ligands can be selected from a group comprising antibodies, antigens, peptides, proteins or glycoproteins, hormones, enzymes, co-factors thereof, substrates or inhibitors of these, polysaccharides, lectins, toxins or anti-toxins, nucleic acids or polynucleotides, haptens or hapten ligands, pigments or dyes.
- the present invention relates to the use of such functionalized foams as modules for bio-purification by affinity in vitro, ex vivo or in vivo of biological molecules or macromolecules.
- the size and geometry of the communicating cavities of the biopolymer foams can be chosen as a function of the cells placed in culture and of their organization in the communicating cavities, more particularly when these cells differentiate within the foam itself.
- the field of cell cultures has experienced significant growth for a very long time, and many devices and products have been developed with the aim of optimizing vital conditions of cells in culture.
- Petrie dish E.g. Petrie dish, CO 2 oven, range of nutrient media, background box treated with products of biological, organic or mineral origin, allowing the cells a better organization, adhesion, proliferation etc. during their cultivation.
- the biopolymer foam defined by the present invention allows the cells cultured, to organize in its communicating cavities, to proliferate and to build transportable and transplantable cellular tissue with or without immunoprotection as will be explained in examples, in particular in transplantation of the neocartilaginous tissue produced by cultured chondrocytes.
- the polymer of the foams of the invention will advantageously include animal or plant cells, in a medium suitable for their proliferation and / or their differentiation.
- One of the first applications of the present invention is the use of this type of foam for the culture of animal or plant cells, where appropriate recombinant for their culture in vitro and the production of biological macromolecules of interest.
- the biocompatible polymer foams according to the invention and more particularly the hydrogel foams, and even more particularly the AN 69 foams find a particularly advantageous application when they contain cells intended for implantations in the human or animal body.
- the advantage of the structure of foams with communicating cavities is that when they are seeded with stem or undifferentiated cells, it is possible to build a cellular tissue inside this foam by preculture in a medium containing the appropriate growth and / or differentiation factors.
- Another type of foam according to the invention carries chondrocytes or chondrogenic stromal cells.
- the implantation of foam carrying chondrocytes allows the production of bio-artificial cartilage or the filling of a bone deficit.
- One way of making these chondrocyte-carrying foams is to separate the chondrocytes from articular cartilage taken from a joint in a human or animal, to seed the chondrocytes in the foam with communicating cavities, to cultivate these chondrocytes seeded in the support immersed in the nutritive medium in a study at 37 ° under an atmosphere comprising 5% of CO 2, and to transplant after culture the foam carrying cells having thus proliferated in articular cartilage in an individual.
- This transplantation can be autologous or heterologous, namely that the chondrocytes can come from an individual donor having tissue compatibility with the recipient (allogenic transplant), or be taken from an individual, cultivated and implanted in the form of foam carrying chondrocytes to level of cartilage or bone to be repaired by the same individual (autologous transplant).
- the foams according to the invention can be seeded with stem or progenitor cells of a particular cell line.
- the marrow is composed of hematopoietic cells in close association with cells of non-hematopoietic origin and a support called the spinal microenvironment.
- stromal cells which are cellular progenitors with multipotent differentiation characteristics. to specific connective tissues such as bone and cartilage.
- Stroma and bone marrow cells which make up around 3% of mononuclear cells, can be isolated by incubating mononuclear cells with coupled endogline (CD15) monoclonal antibodies to magnetic beads.
- CD15 coupled endogline
- This antigen is found in a highly homogeneous cell population with expansion capacities and chondrogenic properties.
- the cell suspension can then be isolated by any means known to those skilled in the art, an example of which can be an affinity column attached to a magnet in order to retain the positive cells which will be collected, analyzed and cultured for expansion.
- This cultivation in the foams according to the invention is carried out in the presence of a culture medium in the presence of appropriate differentiating factors, in particular TGF ⁇ 3. Culture under these conditions thus makes it possible to obtain a pseudo-cellular tissue implantable on the bone or on the cartilage.
- the present invention relates to a foam of biopolymers with communicating cavities carrying hepatocytes. These foams can then be implanted, for example in the peritoneal cavity.
- This transplantation of syngeneic or congenic hepatocytes can make it possible to correct long-term metabolic deficiencies without incurring immunosuppression.
- Such an example on the therapeutic potential of hepatocyte transplantation can be given in N. Gomez et al. (12).
- the transplantation of biocompatible polymer foams, and more particularly still of an AN 69 hydrogel, carrying hepatocytes makes it possible to increase the longevity and the tolerability of the transplant.
- Such a film or such a membrane may advantageously be a hydrogel of the polymer or of the copolymer according to the invention.
- polymer foams according to the invention carrying islets of Langherans.
- the islets of Langherans can be obtained by any technique accessible to those skilled in the art at the time of its implementation. As example, we can cite the technique described in C. Delauney et al (13).
- the transplant carrying the islets of Langherans can thus be assimilated to a bio-artificial pancreas which allows, after implantation, the long-term production of insulin and the regulation of glycemia.
- the foams of biocompatible polymers, and more particularly of hydrogel 69 can constitute cellular reactors implantable in vivo for the production of substances of therapeutic interest.
- the implant carrying the producing cells can be implanted, either subcutaneously, or at the level of a particular organ or tissue.
- we can thus treat different chronic pathologies with therapeutic proteins such as anemia with erythropoietin, hemophilia with factor VIII or factor IX, vascular deficits with angiogenic factors, or solid tumors with anti-angiogenic factors.
- therapeutic proteins such as anemia with erythropoietin, hemophilia with factor VIII or factor IX, vascular deficits with angiogenic factors, or solid tumors with anti-angiogenic factors.
- the feasibility of such implantation techniques has already been shown by E. Payen et al. for erythropoietin (14).
- a mini bio-reactor according to the invention can also contain cells which produce vectors, viruses or recombinant plasmids for gene therapy.
- the bio-reactor can thus be implanted in situ, in particular near the cells which it is desired to treat by this method.
- Another aspect of the invention relates to the use of biocompatible polymer foams according to the invention in the manufacture of a prosthesis intended to fill a deficit of substances in an organ, in particular a breast prosthesis or the complement of a tissue bony.
- the implanted polymer foam does not comprise cells, but comprises a medium allowing cells in contact with said foam to colonize the latter in situ.
- the cells of the organ in which the foam, carrying an appropriate sterile culture medium, is implanted can proliferate and help fill the organ deficit.
- biocompatible polymer foams according to the invention in the manufacture of medicaments for the controlled release of active principle.
- these hydrogel foams offer a particularly suitable means for administering molecular or macromolecular active principles and in particular active principles of peptide or polypeptide nature.
- the three-dimensional foams with communicating cells according to the invention will find their application each time a person skilled in the art wishes to produce an implant with undifferentiated or differentiated cells of a certain given type.
- the above examples are not limiting insofar as in the same way one can envisage an implant carrying neuronal cells, keratinocytes, etc.
- foams will also find their application each time that one wishes to carry out before implantation an in situ differentiation of stem cells, constituting the frame of a preformed tissue which can then be grafted advantageously on an organ or a tissue which one wishes to fill a deficient function.
- Figure 1 photograph taken under optical microscopy of a basal bone regrowth and articular cartilaginous surface in the porous material with three-dimensional structure previously sown with rabbit chondrocytes and implanted on the femoral condyle after creation of a cartilaginous deficit .
- Figure 1a is a photo at 15 times magnification and
- Figure 1b is a photo at 60 times magnification of the central part of photograph 1a showing the detail of the cartilaginous regrowth.
- Figure 2 photograph of an AN 69 polymer foam obtained by the method of Example 4 below.
- the top photo (2a) is at x 17 magnification and the bottom photo (2b) is at x 100 magnification.
- Example 1 Realization of a flexible foam with communicating cavities, before the approximate dimensions of 0.8 cm x 1.3 cm x 2.1 cm and the size of the pores of a few tenths of a millimeter
- the polymer solution consisting of:
- aqueous sodium chloride solution at 9 g / l; - 91% of dimethylsulfoxide (DMSO) by successive dissolution of the components with stirring and at 50 ° C.
- DMSO dimethylsulfoxide
- TPX polymethylpentene
- Example 2 Production of a semi-rigid foam with communicating cavities carrying chondrocytes
- a silicone elastomer mold is prepared, which has a flat and smooth bottom, bordered in turn by a bead 1.5 mm thick.
- porous elastomer plate is then decontaminated with a solution containing peracetic acid (APA), carefully washed with sterile physiological serum until the last traces of APA disappear. Then, it is sown by the chondrocytes isolated from the articular cartilage of the rabbit, either by their injection into the syringe provided with a equilla, or by "infiltration / aspiration" as in a sponge. Placed in a Petri dish containing a nutritive liquid, the porous elastomer plate with communicating cells seeded with chondrocytes is subjected to a cell culture process in a CO 2 study.
- APA peracetic acid
- Example 3 Production of a porous material of small thickness (0.5 mm), as a support for living cells, in particular for keratocytes
- Example 4 Production of a polymer filter block with communicating cells, which can be used for various treatments of the fluids circulating in pores through this filter block
- Dimethylformamide In a glass or plastic beaker resistant to Dimethylformamide, 9% of this solution are mixed with a spatula with 91% of crystallized cane sugar. This mixture is then transferred to another glass or plastic beaker resistant to Dimethylformamide, and this mixture is packed using a curved spatula and / or using a cylinder or another Beccher, having a slightly smaller diameter, so that it can function as a compacting piston.
- the beaker is immersed with the packed mixture in distilled water or in an aqueous solution composed of various mineral or organic salts, preferably biologically acceptable. After a few minutes, the filter block thus formed is removed from the mold, and the sugar crystals are allowed to dissolve by continuous or batch washing with distilled water or with aqueous solutions of the various salts.
- Example 5 Obtaining a form of polymer with communicating cells by molding the mixture of the polymer solution and the crystallized sugar
- Example 4 An identical mixture is therefore prepared as described in Example 4. This mixture is then introduced with a spatula and packed using a polytetrafluoroethylene rod in a glass tube. Distilled water is then introduced into the interior of the tube and a cylinder is released by gravity or by a weak stream of water, which after dissolution total of crystallized sugar becomes a polymer cylinder with communicating cells.
- Example 6 Production of a polymer block AN 69 containing in communicating cells iusgu 'with 97% water and which can be used after necessary examinations as an implant for filling the mango substance (breast implant for example)
- a mixture is prepared containing 3.5% of the polymer solution (25% copolymer of acrylonitrile and sodium methallylsulfonate and 75% of Dimethylformamide) and 96.5% of the crystallized sugar. This mixture is transferred to a mold, it is packed, then the filled mold is immersed in water or in aqueous solutions of the various salts. After the sugar has completely dissolved, a block of porous elastomer is obtained, containing almost 97% of water in its cavities.
- Example 7 Production of a porous material with communicating cavities in Polyethersulfone
- Example 8 Production of a porous material with communicating cavities in Povhvdroxypropyl methacrylate
- Example 10 Production of a rigid and porous material with tight communicating cavities in Polysulfone
- Collier C "Preliminary report on cell encapsulation in a hydrogel made of biocompatible material, AN 69, for the development of a bioartificial pancreas", The International Journal of Artificial Organs (1994), vol. 17, 1: 046-052. (8) Honiger J., Balladur P., Mariani P., Calmus Y., Vaubourdolle M.,
Description
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Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
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FR0008186A FR2810552B1 (fr) | 2000-06-26 | 2000-06-26 | Polymere biocompatible a structure tridimensionnelle a cellules communicantes, procede de preparation et application en medecine et en chirurgie |
FR0008186 | 2000-06-26 | ||
PCT/FR2001/002007 WO2002000775A1 (fr) | 2000-06-26 | 2001-06-25 | Polymere biocompatible a structure tridimensionnelle a cellules communicantes, procede de preparation et application en medecine et en chirurgie |
Publications (2)
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EP1412421A1 true EP1412421A1 (fr) | 2004-04-28 |
EP1412421B1 EP1412421B1 (fr) | 2012-03-07 |
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EP01949519A Expired - Lifetime EP1412421B1 (fr) | 2000-06-26 | 2001-06-25 | Polymere biocompatible a structure tridimensionnelle a cellules communicantes, procede de preparation et application en medecine et en chirurgie |
Country Status (8)
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US (1) | US20040062809A1 (fr) |
EP (1) | EP1412421B1 (fr) |
JP (1) | JP2004501700A (fr) |
AT (1) | ATE548414T1 (fr) |
AU (1) | AU2001270656A1 (fr) |
CA (1) | CA2414521A1 (fr) |
FR (1) | FR2810552B1 (fr) |
WO (1) | WO2002000775A1 (fr) |
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UA84862C2 (en) * | 2003-03-03 | 2008-12-10 | Месье-Бугатти | Substrate |
US7713542B2 (en) | 2005-01-14 | 2010-05-11 | Ada Foundation | Three dimensional cell protector/pore architecture formation for bone and tissue constructs |
US9671301B2 (en) * | 2006-09-29 | 2017-06-06 | Wake Forest University Health Sciences | Small-scale pressure sensors |
DE102007024642A1 (de) * | 2007-05-24 | 2008-11-27 | Eyesense Ag | Hydrogel-Implantat für Sensorik von Metaboliten am Auge |
EP2314672B1 (fr) | 2008-09-25 | 2015-04-15 | Gambro Lundia AB | Rein bio-artificiel hybride |
EP2168668A1 (fr) | 2008-09-25 | 2010-03-31 | Gambro Lundia AB | Membrane pour expansion cellulaire |
EP2177603A1 (fr) | 2008-09-25 | 2010-04-21 | Gambro Lundia AB | Dispositif pour expansion cellulaire rénale |
EP2168666A1 (fr) | 2008-09-25 | 2010-03-31 | Gambro Lundia AB | Membrane irradiée pour expansion cellulaire |
FR2991988B1 (fr) * | 2012-06-15 | 2015-08-07 | Laurent Laroche | Procede de preparation d'objets en hydrogel biocompatible pour leur application dans le domaine medical, et plus particulierement en ophtalmologie |
US20160200891A1 (en) * | 2013-08-22 | 2016-07-14 | Polyvalor Limited Partnership | Porous gels and methods for their preparation |
WO2016148246A1 (fr) | 2015-03-18 | 2016-09-22 | 富士フイルム株式会社 | Matériau pour la régénération du cartilage et procédé pour le produire |
JP6586160B2 (ja) * | 2015-03-18 | 2019-10-02 | 富士フイルム株式会社 | 軟骨再生材料 |
FR3034307B1 (fr) | 2015-04-03 | 2021-10-22 | Univ Grenoble 1 | Reacteur intestinal implantable |
CN104840272B (zh) * | 2015-05-11 | 2016-09-07 | 浙江大学 | 一种具有内置营养通道的三维生物结构的打印方法 |
CA3023221A1 (fr) | 2016-05-05 | 2017-11-09 | Southwest Research Institute | Bioreacteur tridimensionnel pour expansion cellulaire et applications associees |
US11149244B2 (en) | 2018-04-04 | 2021-10-19 | Southwest Research Institute | Three-dimensional bioreactor for T-cell activation and expansion for immunotherapy |
US10410542B1 (en) | 2018-07-18 | 2019-09-10 | Simulated Inanimate Models, LLC | Surgical training apparatus, methods and systems |
WO2020068840A1 (fr) | 2018-09-24 | 2020-04-02 | Southwest Research Institute | Bioréacteurs tridimensionnels |
US11492580B2 (en) | 2020-05-12 | 2022-11-08 | Southwest Research Institute | Method using a three-dimensional bioprocessor |
Family Cites Families (6)
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GB8708476D0 (en) * | 1987-04-09 | 1987-05-13 | Charlesworth D | Making polymer material |
US5273750A (en) * | 1988-05-02 | 1993-12-28 | Institute National De La Sante Et De La Recherche Medicale- Inserm | Uncrosslinked hydrogel, process for its preparation and its uses as an article for medical and/or surgical purposes such as tubes, films, joints, implants and the like, particularly in ophthalmology |
CA2049377A1 (fr) * | 1990-09-26 | 1992-03-27 | Donald V. Hillegass | Implant corporel biocompatible a surface texturee |
US5514378A (en) * | 1993-02-01 | 1996-05-07 | Massachusetts Institute Of Technology | Biocompatible polymer membranes and methods of preparation of three dimensional membrane structures |
US5626861A (en) * | 1994-04-01 | 1997-05-06 | Massachusetts Institute Of Technology | Polymeric-hydroxyapatite bone composite |
US6268405B1 (en) * | 1999-05-04 | 2001-07-31 | Porex Surgical, Inc. | Hydrogels and methods of making and using same |
-
2000
- 2000-06-26 FR FR0008186A patent/FR2810552B1/fr not_active Expired - Fee Related
-
2001
- 2001-06-25 AT AT01949519T patent/ATE548414T1/de active
- 2001-06-25 JP JP2002505895A patent/JP2004501700A/ja active Pending
- 2001-06-25 CA CA002414521A patent/CA2414521A1/fr not_active Abandoned
- 2001-06-25 EP EP01949519A patent/EP1412421B1/fr not_active Expired - Lifetime
- 2001-06-25 WO PCT/FR2001/002007 patent/WO2002000775A1/fr active Application Filing
- 2001-06-25 AU AU2001270656A patent/AU2001270656A1/en not_active Abandoned
-
2002
- 2002-12-26 US US10/329,651 patent/US20040062809A1/en not_active Abandoned
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Title |
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See references of WO0200775A1 * |
Also Published As
Publication number | Publication date |
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FR2810552A1 (fr) | 2001-12-28 |
ATE548414T1 (de) | 2012-03-15 |
AU2001270656A1 (en) | 2002-01-08 |
JP2004501700A (ja) | 2004-01-22 |
CA2414521A1 (fr) | 2002-01-03 |
FR2810552B1 (fr) | 2004-11-19 |
US20040062809A1 (en) | 2004-04-01 |
WO2002000775A1 (fr) | 2002-01-03 |
EP1412421B1 (fr) | 2012-03-07 |
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